Figure 3

Scheme of selection strategy leading to TAA identification. A phage-displayed tumor cDNA library is preincubated with patient serum. TAA-specific antibodies bind to antigens exposed on the phage surface. Abs-phage complex is captured by protein A-coated solid support (ELISA plate or dynebeads). Non-bound phage are washed away. Bound phage are eluted by infection of added bacteria and amplified. Positive clones are isolated by immunoscreening procedure and then picked in ordered array on a bacterial lawn, transferred to nitrocellulose membrane and probed with different positive and negative sera.